The HIV pandemic has caused an estimated 33 million deaths to date. In the U.S. alone, 1.2 million people currently live with HIV and 34,800 new cases were diagnosed in 2019. This adds up to an estimated $16.4 billion for a lifetime of HIV treatment for patients in the U.S. HIV infects immune cells which express the CD4 and CXCR4/CCR5 co-receptors including helper T-cells. Even after a patient mounts an initial, effective immune response, the virus persists undetected by the immune system in quiescently infected CD4+ T-cells. For this reason, the primary treatment for HIV, combined antiretroviral therapy (cART), is not able to eradicate HIV from patients. Many HIV patients are able to live with continuous treatment, but this is both costly and uncertain to remain effective given the recent identification of a more virulent strain of HIV-1 in which patients suffer an accelerated loss of CD4+ T-cells. There is a critical need for innovative, efficacious therapeutics to direct a more robust immune attack of the virus. One promising strategy is to generate more active anti-HIV CD8+ cells. Chimeric Antigen Receptor-T cell (CAR- T) is a relatively new process in which patient T-cells can be programmed to attack cells expressing a target protein characteristic of a specific disease. When used with agents to make infected cells visible, it has been hypothesized that CAR-T will support the host immune system to fully eradicate HIV-infected cells. There are currently several technological limitations to the production of CAR-T cells, and the rate limiting step is transfer of genetic material into the primary T-cells to program them to eliminate cells expressing a target protein. Current methods of creating CAR-T cells demonstrate an inefficiency barring translation from the laboratory to clinical settings. Lipofection inefficiently transfects primary T-cells, and electroporation and biolistics both damage cells. Retroviruses are limited to 8-10 kb of genetic material which limits advanced applications, are toxic to cells if used at too high a dose, and are complex to construct. In addition, retroviruses integrate into their target cell's genome which could inactivate a tumor suppressor gene and create tumors. AGTC has developed a novel method of introducing biomolecules into mammalian cells using an array of closely packed, aligned carbon nanotubes to achieve highly efficient transfer with low cytotoxicity and high capacity for genetic cargo. This technology has potential to overcome size limits of current gene-transfer technologies in addition to being simpler, faster, and more flexible. In this proposal, AGTC will (1) optimize gene transfer into primary human T-cells using carbon nanotube technology (CNT); and (2) create human anti-HIV CD8+ CAR-T cells against HIV envelope glycoprotein, gp120. Successful completion of the proposed work will enhance knowledge about CNT capabilities, and allow AGTC to proceed to further testing of the CAR-T cell product with an eye to therapeutic use in HIV positive patients.
Public Health Relevance Statement: NARRATIVE About 1.2 million people in the U.S. live with HIV which generates a health care burden of about $16.4 billion for a lifetime of treatment. The primary treatment for HIV is combined antiretroviral therapy (cART); however, despite being able to slow or stop HIV infection, cART is not able to completely eradicate HIV from patients. The current proposal will create Chimeric Antigen Receptor-T cells (CAR-T), a relatively new method to enhance the immune response, using our proprietary carbon nanotube technology (CNT) to target and eliminate HIV infected cells.
Project Terms: Acceleration; inhibitor; Biological Assay; Assay; Bioassay; Biologic Assays; Cell Survival; Cell Viability; Cells; Cell Body; Cessation of life; Death; Diagnosis; Disease; Disorder; DNA; Deoxyribonucleic Acid; Eye; Eyeball; Flow Cytometry; Flow Cytofluorometries; Flow Cytofluorometry; Flow Microfluorimetry; Flow Microfluorometry; flow cytophotometry; Future; Genome; Goals; Helper-Inducer T-Lymphocyte; Helper Cells; Helper T-Cells; Helper T-Lymphocytes; Helper-Inducer T-Cells; Inducer Cells; Inducer T-Lymphocytes; HIV; AIDS Virus; Acquired Immune Deficiency Syndrome Virus; Acquired Immunodeficiency Syndrome Virus; Human Immunodeficiency Viruses; LAV-HTLV-III; Lymphadenopathy-Associated Virus; Virus-HIV; HIV Envelope Protein gp120; HIV Envelope Glycoprotein gp120; HIV env Protein gp120; HTLV-III gp120; gp120; gp120 ENV Glycoprotein; gp120(HIV); HIV Infections; HTLV-III Infections; HTLV-III-LAV Infections; Human T-Lymphotropic Virus Type III Infections; HIV Seropositivity; Anti-HIV Positivity; HIV Positive; HIV Positivity; HIV Seroconversion; HIV antibody positive; HTLV-III Seroconversion; HTLV-III Seropositivity; HIV-1; HIV-I; HIV1; Human Immunodeficiency Virus Type 1; Human immunodeficiency virus 1; Human; Modern Man; Immune system; Immunity; Laboratories; Lactate Dehydrogenase; EC 1.1.1.27; L-Lactate Dehydrogenase; L-Lactic Acid Dehydrogenase; NAD-Lactate Dehydrogenase; lactic acid dehydrogenase; Methods; Fluorescence Microscopy; Fluorescence Light Microscopy; Persons; Patients; Plasmids; Production; Proteins; Retroviridae; Retroviruses; Virus-Retrovirus; T-Lymphocyte; T-Cells; thymus derived lymphocyte; Cytotoxic T-Lymphocytes; Cell-Mediated Lympholytic Cells; Cytolytic T-Cell; Cytotoxic T Cell; killer T cell; CD4 Positive T Lymphocytes; CD4 Cells; CD4 T cells; CD4 helper T cell; CD4 lymphocyte; CD4+ T-Lymphocyte; CD4-Positive Lymphocytes; T4 Cells; T4 Lymphocytes; Technology; Testing; Transfection; Translations; translation; Virus Diseases; Viral Diseases; viral infection; virus infection; virus-induced disease; Virus; Work; Tumor Suppressor Genes; Anti-Oncogenes; Antioncogenes; Cancer Suppressor Genes; Emerogenes; Onco-Suppressor Genes; Oncogenes-Tumor Suppressors; Recessive Oncogenes; Tumor Suppressing Genes; oncosuppressor gene; health care; Healthcare; Mediating; bases; base; Surface; Clinical; Phase; electroporative delivery; gene electrotransfer; Electroporation; Chemicals; CD8 Cell; CD8 T cells; CD8 lymphocyte; CD8+ T cell; CD8+ T-Lymphocyte; CD8-Positive Lymphocytes; T8 Cells; T8 Lymphocytes; CD8-Positive T-Lymphocytes; Technology Transfer; uptake; Co-culture; Cocultivation; Coculture; Coculture Techniques; Immunological response; host response; immune system response; immunoresponse; Immune response; cell mediated therapies; cell-based therapeutic; cell-based therapy; cellular therapeutic; cellular therapy; Cell Therapy; Therapeutic; Genetic; Deposition; Deposit; Jurkat Cells; Knowledge; programs; Immune; Immunes; Complex; Genetic Materials; cytotoxicity; Lytotoxicity; receptor; Receptor Protein; vapor; cell injury; Cellular injury; cell damage; cellular damage; damage to cells; injury to cells; cell killing; novel; Devices; rapid technique; rapid method; gene gun; Biolistics; Gene-Gun Technique; HIV therapy; Buckytubes; Carbon nano tubes; Carbon Nanotubes; nano tubes; Nanotubes; Therapeutic Uses; EGFP protein; enhanced green fluorescent protein; Effectiveness; AIDS test; AIDS/HIV test; HIV test; HIV-1 test; HIV-2 test; Human immunodeficiency virus test; CCR5 gene; C-C CKR-5; C-C CKR-5 Gene; C-C Chemokine Receptor Type 5; C-C Chemokine Receptor Type 5 Gene; CC Chemokine Receptor 5; CC-CKR-5; CC-CKR-5 Gene; CC-CKR5; CCCKR5; CCCKR5 Gene; CCR-5; CCR-5 Gene; CCR5; CCR5 Protein; CCR5 Receptors; CD195 Antigen; CD195 Antigen Gene; CHEMR13; CHEMR13 Gene; CKR-5; CKR-5 Gene; CKR5; CKR5 Gene; CKR5 Receptors; CMKBR5; CMKBR5 Gene; Chemokine (C-C Motif) Receptor 5; Chemokine (C-C) Receptor 5; Chemokine (C-C) Receptor 5 Gene; HIV-1 Fusion Co-Receptor; HIV-1 Fusion Co-Receptor Gene; CXCR4 gene; CXC-R4; CXCR-4; CXCR4; D2S201E; FB22; HM89; HSY3RR; LAP3; LCR1; LESTR; NPY3R; NPYR; NPYRL; NPYY3R; CD8B1 gene; CD8; CD8B; CD8B1; LYT3; Length; Dose; Mammalian Cell; Proliferating; in vivo; Gene Transfer; Virulent; Cellular Morphology; cell morphology; Characteristics; Process; Development; developmental; pandemic disease; pandemic; Nanomanufacturing; antiretroviral therapy; anti-retroviral therapy; anti-retroviral treatment; antiretroviral treatment; care burden; cost; manufacturing process; innovate; innovative; innovation; lipofection; murine model; mouse model; plasmid DNA; tumor; flexible; flexibility; chimeric antigen T cell receptor; chimeric antigen receptor; Geometry; humanized mice; humanized mouse; Virus reservoir; Viral reservoir; experiment; experimental research; experiments; experimental study; CAR T cells; CAR modified T cells; CAR-T; CAR-Ts; T cells for CAR; chimeric antigen receptor (CAR) T cells; chimeric antigen receptor fusion protein T-cells; chimeric antigen receptor modified T cells; chimeric antigen receptor T cells; CAR T therapy; chimeric antigen receptor (CAR) T cell therapy; chimeric antigen receptor T cell therapy; CAR T cell therapy; Diameter; cost comparison; compare cost; Good Manufacturing Process; Good manufacturing practice; manufacture
